A major challenge in the research on membrane protein is to deal with the absence of the lipid bilayer environment, which requires a mimic “native-like” lipid bilayer. Nanodisc, using amphiphilic membrane scaffold proteins (MSPs) to stabilize phospholipid molecules in the aqueous phase, self-assembly forms a disc-like structure similar to the natural phospholipid bilayer membrane environment. As a cutting-edge technique, nanodiscs are reported to provide model membrane environment of water solubility, homogeneity, control of oligomerization state and size, composition and specific functional modification on nanometer scale as well. In this review, we introduced the reconstitution of MSPs and integral proteins to introduce nanodiscs, followed by the nanodiscs applications in structural, functional and medical fields of membrane proteins. As for its medical applications, the high efficiency of delivery capacity as a carrier of hydrophobic drugs, and the targeting specificity of antineoplastic were particularly highlighted. Nanodisc technique also help to throw a light on the structural resolution and functional characterization of membrane proteins in imaging technology. Moreover, nanodiscs are applied to cardiovascular disease to boost the efficiency and manoeuvrability of cholesterol transport. In summary, nanodisc technique provides new method and insight into membrane protein research in the future, for better diagnosis and treatment of clinical diseases.